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Anal Biochem. 2011 Feb 1;409(1):37-45. doi: 10.1016/j.ab.2010.09.040. Epub 2010 Oct 23.

Comparison of metalloproteinase protein and activity profiling.

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  • 1Department of Chemistry and Biochemistry, Florida Atlantic University, Boca Raton, FL 33431, USA.

Abstract

Proteolytic enzymes play fundamental roles in many biological processes. Members of the matrix metalloproteinase (MMP) family have been shown to take part in processes crucial in disease progression. The current study used the ExcelArray Human MMP/TIMP Array to quantify MMP and tissue inhibitor of metalloproteinase (TIMP) production in the lysates and media of 14 cancer cell lines and 1 normal cell line. The overall patterns were very similar in terms of which MMPs and TIMPs were secreted in the media versus associated with the cells in the individual samples. However, more MMP was found in the media (in both amount and variety). TIMP-1 was produced in all cell lines. MMP activity assays with three different fluorescence resonance energy transfer (FRET) substrates were then used to determine whether protein production correlated with function for the WM-266-4 and BJ cell lines. Metalloproteinase activity was observed for both cell lines with a general MMP substrate (Knight SSP), consistent with protein production data. However, although both cell lines promoted the hydrolysis of a more selective MMP substrate (NFF-3), metalloproteinase activity was confirmed only in the BJ cell line. The use of inhibitors to confirm metalloproteinase activities pointed to the strengths and weaknesses of in situ FRET substrate assays.

Copyright © 2010 Elsevier Inc. All rights reserved.

PMID:
20920458
[PubMed - indexed for MEDLINE]
PMCID:
PMC3298814
Free PMC Article

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